U.S. patent number 11,136,336 [Application Number 17/071,081] was granted by the patent office on 2021-10-05 for thienopyrimidones.
This patent grant is currently assigned to Boehringer Ingelheim International GmbH. The grantee listed for this patent is Boehringer Ingelheim International GmbH. Invention is credited to Florian Paul Christian Binder, Georg Dahmann, Martin Thomas Fleck, Joerg P. Hehn, Annekatrin Charlotte Heimann, Jens Willwacher.
United States Patent |
11,136,336 |
Fleck , et al. |
October 5, 2021 |
Thienopyrimidones
Abstract
This invention relates to thienopyrimidinones and their use as
inhibitors of TRPA1 activity, pharmaceutical compositions
containing the same, and methods of using the same as agents for
treatment and/or prevention of fibrotic diseases, inflammatory and
auto-immune diseases and CNS-related diseases.
Inventors: |
Fleck; Martin Thomas (Munich,
DE), Binder; Florian Paul Christian (Maselheim,
DE), Dahmann; Georg (Biberach, DE), Hehn;
Joerg P. (Biberach an der Riss, DE), Heimann;
Annekatrin Charlotte (Biberach an der Riss, DE),
Willwacher; Jens (Biberach an der Riss, DE) |
Applicant: |
Name |
City |
State |
Country |
Type |
Boehringer Ingelheim International GmbH |
Ingelheim am Rhein |
N/A |
DE |
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Assignee: |
Boehringer Ingelheim International
GmbH (Ingelheim am Rhein, DE)
|
Family
ID: |
68289872 |
Appl.
No.: |
17/071,081 |
Filed: |
October 15, 2020 |
Prior Publication Data
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Document
Identifier |
Publication Date |
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US 20210107918 A1 |
Apr 15, 2021 |
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Foreign Application Priority Data
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Oct 15, 2019 [EP] |
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19203171 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
C07D
495/04 (20130101); C07D 495/14 (20130101); A61P
11/00 (20180101) |
Current International
Class: |
C07D
495/04 (20060101); C07D 495/14 (20060101) |
Foreign Patent Documents
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2012085662 |
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Jun 2012 |
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WO |
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2012172475 |
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Dec 2012 |
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WO |
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2016023832 |
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Feb 2016 |
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WO |
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2016133888 |
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Aug 2016 |
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WO |
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2017060488 |
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Apr 2017 |
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WO |
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Other References
Borate et al., "Synthesis and evaluation of
thieno[2,3-d]pyrimidin-4(3H)-ones as potential antitubercular
agents", Medchemcomm, vol. 6, No. 12, Jan. 1, pp. 2209-2215, 2013.
cited by applicant .
El-Tombary et al., "Synthesi s and 1-13 Biological Evaluation of
Some Novel Thieno[2,3-d] pyrimidine Derivatives as Potential
Anti-inflammatory and Analgesie Agents", Medicinal Chemistry, vol.
9, No. 8, Oct. 1, 2013, pp. 1099-1112. cited by applicant .
International Search Report for corresponding application,
PCT/EP2020/078853, dated Nov. 6, 2020. cited by applicant .
Schenkel et al., "11 Optimization of a Novel Quinazoline-Based
Series of Transient Receptor Potential AI (TRPAI) Antagonists
Demonstrating Potent in Vivo Activity 11", J . Med. Ch Em ., 2016,
vol. 59, pp. 2794-28. cited by applicant.
|
Primary Examiner: Anderson; Rebecca L
Attorney, Agent or Firm: Datlow; Philip I.
Claims
The invention claimed is:
1. A compound according to formula (I) ##STR00082## wherein A is
selected from the group consisting of phenyl, naphthyl, thiophenyl,
benzothiophenyl and benzofuranyl, each optionally substituted with
one or two members selected the group consisting of H, F, Cl, Br,
C.sub.1-4-alkyl, F.sub.1-3-fluoro-C.sub.1-4-alkyl, CN, OCH.sub.3,
cyclopropyl, and cyclobutyl, or A is selected from ##STR00083## and
R.sup.1 is selected from H, C.sub.1-4-alkyl,
F.sub.1_3-fluoro-C.sub.1-4-alkyl, C.sub.1-4-alkyl-OH or
C.sub.1-4-alkyl-CN; R.sup.2 is selected from C.sub.1-2-alkyl or Cl;
or R.sup.1 and R.sup.2 are each CH.sub.2 joined via a bond forming
a 6-membered ring; or a pharmaceutically acceptable salt
thereof.
2. The compound of formula (I) according to claim 1, wherein A is
selected from the group consisting of phenyl and benzofuranyl, each
optionally substituted with one or two members selected from the
group consisting of H, F, Cl, Br, C.sub.1-4-alkyl,
F.sub.1-3-fluoro-C.sub.1-4-alkyl, CN, OCH.sub.3, cyclopropyl, and
cyclobutyl, or A is selected from ##STR00084## or a
pharmaceutically acceptable salt thereof.
3. The compound of formula (I) according to claim 1, wherein A is
selected from the group consisting of phenyl and benzofuranyl, each
optionally substituted with one or two members selected from the
group consisting of H, F, Cl, and CH.sub.3; or A is selected from
##STR00085## or a pharmaceutically acceptable salt thereof.
4. The compound of formula (I) according to claim 1, wherein A is
selected from the group consisting of ##STR00086## or a
pharmaceutically acceptable salt thereof.
5. The compound of formula (I) according to claim 1, wherein
R.sup.1 is selected from the group consisting of H, CH.sub.3,
CH.sub.2CHF.sub.2 and CH.sub.2C(CH.sub.3).sub.2OH; or a
pharmaceutically acceptable salt thereof.
6. The compound of formula (I) according to claim 1, wherein
R.sup.2 is CH.sub.3 or Cl; or a pharmaceutically acceptable salt
thereof.
7. The compound of formula (I) according to claim 1, selected from
the group consisting of: ##STR00087## or a pharmaceutically
acceptable salt thereof.
8. The (S)-enantiomer of the compound of formula (I) according to
claim 1, selected from the group consisting of ##STR00088##
##STR00089## or a pharmaceutically acceptable salt thereof.
9. A pharmaceutically acceptable salt of a compound according to
claim 1.
10. A pharmaceutical composition comprising at least one compound
according to claim 1, or a pharmaceutically acceptable salt
thereof, and one or more pharmaceutically acceptable
excipients.
11. A method for treating or preventing an inflammatory airway
disease, a fibrotic disease or cough comprising administering to a
patient having such disease or cough an effective amount of a
compound according to claim 1 or a pharmaceutically acceptable salt
thereof.
12. A method according to claim 11, wherein the patient has
idiopathic pulmonary fibrosis (IPF) or cough.
Description
FIELD OF THE INVENTION
This invention relates to thienopyrimidinones and their use as
inhibitors of TRPA1 activity, pharmaceutical compositions
containing the same, and methods of using the same as agents for
treatment and/or prevention of fibrotic diseases, inflammatory and
auto-immune diseases and CNS-related diseases.
BACKGROUND INFORMATION
Transient receptor potential channels (TRP channels) are a group of
voltage-gated ion channels located mostly on the plasma membrane of
numerous mammalian cell types. There are approximately 30
structurally related TRP channels sorted into groups: TRPA, TRPC,
TRPM, TRPML, TRPN, TRPP and TRPV. Transient receptor potential
cation channel, subfamily A, member 1 (TRPA1), also known as
transient receptor potential ankyrin 1, is the only member of the
TRPA gene subfamily. Structurally, TRPA channels are characterized
by multiple N-terminal ankyrin repeats (.about.14 in the N-terminus
of human TRPA1) that gives rise to the "A" for ankyrin designation
(Montell, 2005).
TRPA1 is highly expressed in the plasma membrane of sensory
neutrons in the dorsal root and nodose ganglia that serve both skin
and lung, as well as in small intestine, colon, pancreas, skeletal
muscle, heart, brain, bladder and lymphocytes
(https://www.proteinatlas.org/) as well as in human lung
fibroblasts.
TRPA1 is best known as a sensor for environmental irritants giving
rise to somatosensory modalities such as pain, cold and itch. TRPA1
is activated by a number of reactive, electrophilic stimuli (e.g.
allyl isothiocyanate, reactive oxygen species), as well as
non-reactive compounds (e.g. icilin), implicated in cough
associated with asthma, chronic pulmonary obstructive disease
(COPD), idiopathic pulmonary fibrosis (IPF) or post-viral cough or
for chronic idiopathic cough as well as cough in sensitive
patients. (Song and Chang, 2015; Grace and Belvisi, 2011). TRPA1
inhibitors are useful in the treatment of IPF in which cough is
highly prevalent because of the link between cough and lung injury,
based on studies showing cough-induced elevation of TGF-.beta. (Xie
et al., 2009; Froese et al., 2016; Tschumperlin et al., 2003;
Yamamoto et al., 2002; Ahamed et al., 2008). TRPA1 antagonists
inhibit calcium signaling triggered by cough triggers such as
cigarette smoke extract (CSE) oxidative stress, inflammatory
mediator release and downregulated antioxidant gene expression (Lin
et al., 2015; Wang et al., 2019). TRPA1 antagonists are effective
in studies of atopic dermatitis (Oh et al., 2013; Wilson et al.,
2013), contact dermatitis (Liu et al., 2013), psoriasis-associated
itch (Wilson et al., 2013) and IL-31-dependent itch (Cevikbas et
al., 2014). A human TRPA1 gain-of-function has been associated with
familial episodic pain syndrome (Kremeyer et al., 2010). A TRPA1
antagonist was effective in a behavioral model of migraine-related
allodynia (Edelmayer et al., 2012). TRPA1 is selectively increased
in trigeminal ganglia innervating injured teeth when compared to
TRPA1 expression in trigeminal ganglia innervating healthy teeth
(Haas et al., 2011). Several anaesthetics are known to be TRPA1
agonists, including isoflurane (Matta et al., 2008) providing
rationale for TRPA1 inhibitors for the relief of post-surgical
pain. TRPA1 knockout mice and wild type mice treated with a TRPA1
antagonist showed anxiolytic- and antidepressant-like phenotypes
(de Moura et al., 2014). TRPA1 inhibitors are expected to have
benefit in the treatment of diabetic neuropathy based on studies
showing a mechanistic link of inverse regulation between AMPK and
TRPA1 (Hiyama et al., 2018; Koivisto and Pertovaara, 2013; Wang et
al., 2018). TRPA1 knockout mice exhibit smaller myocardial infarct
sizes compared to wild type mice (Conklin et al., 2019). TRPA1
knockout and pharmacological intervention inhibited TNBS-induced
colitis in mice (Engel et al., 2011). In a mouse brain ischaemia
model, TRPA1 knock-out and TRPA1 antagonists reduce myelin damage
(Hamilton et al., 2016). Urate crystals and joint inflammation are
reduced in TRPA1 knockout mice in a monosodium urate mouse model of
gout (Moilanen et al., 2015). TRPA1 deletion in rats ameliorated
joint inflammation and hyperalgesia in a rat model of acute gout
flares (Trevisan et al., 2014). Activation of TRPA1 elicits an
inflammatory response in osteoarthritic chondrocytes (Nummenmaa et
al., 2016). TRPA1 inhibition and genetic deletion reduces
inflammatory mediators in osteoarthritic mouse chondrocytes and
murine cartilage (Nummenmaa et al., 2016). Finally, TRPA1 knockout
mice exhibited improvements in weight bearing on the osteoarthritic
limb in an MIA-evoked knee swelling model (Horvath et al., 2016).
TRPA1 is differentially expressed in the in the bladder epithelium
rats (Du et al., 2007) and of patients with bladder outlet
obstruction (Du et al., 2008). TRPA1 receptor modulation attenuates
bladder overactivity in a rat model of spinal cord injury (Andrade
et al., 2011) and intrathecal administration of TRPA1 antagonists
attenuate cyclophosphamide-induced cystitis in rats with
hyper-reflexia micturition (Chen et al., 2016).
It is therefore desirable to provide potent TRPA1 inhibitors.
TRPA1 inhibitors of various structural classes are reviewed in S.
Skerratt, Progress in Medicinal Chemistry, 2017, Volume 56, 81-115
and in D. Preti, G. Saponaro, A. Szallasi, Pharm. Pat. Anal. (2015)
4 (2), 75-94.
WO2017/060488 discloses compounds that are antagonists of TRPA1,
having the generalized structural formula
##STR00001##
The TRPA1 activity of Examples 53, 72, 73, 86 and 90 therein is
disclosed having IC.sub.50's of less than 100 nM in a calcium flux
assay.
L. Schenkel, et al., J. Med. Chem. 2016, 59, 2794-2809 discloses
quinazolinone-based TRPA1 antagonists including compounds of the
generalized structural formula
##STR00002## of which compound 31, wherein R is OH, is disclosed as
having an antagonistic TRPA1 activity of IC.sub.50 58 nM in a FLIPR
assay and having an intrinsic clearance in human liver microsomes
of <14 .mu.L/min/kg.
DETAILED DESCRIPTION OF THE INVENTION
The present invention provides novel thienopyrimidinones that are
surprisingly potent inhibitors of TRPA1 (Assay A), further
characterised by improved stability in human liver microsomes
(Assay B) improved stability in human hepatocytes (Assay C).
Compounds of the present invention differ structurally from
examples 53, 72, 73, 86 and 90 in WO2017/060488 and from example 31
in L. Schenkel, et al., J. Med. Chem. 2016, 59, 2794-2809, in that
they contain a thienopyrimidinone core with amido substituents as
well as substituents adjacent to a secondary aliphatic alcohol.
These structural differences unexpectedly lead to a favourable
combination of (i) inhibition of TRPA1, (ii) stability in human
liver microsomes, and (iii) stability in human hepatocytes.
Compounds of the invention are thus superior to those disclosed in
the prior art in terms of the combination of the following
parameters: potency as inhibitors of TRPA1 stability in human liver
microsomes stability in human hepatocytes
Stability in human liver microsomes refers to the susceptibility of
compounds to biotransformation in the context of selecting and/or
designing drugs with favorable pharmacokinetic properties as a
first screening step. The primary site of metabolism for many drugs
is the liver. Human liver microsomes contain the cytochrome P450s
(CYPs), and thus represent a model system for studying phase I drug
metabolism in vitro. Enhanced stability in human liver microsomes
is associated with several advantages, including increased
bioavailability and adequate half-life, which can enable lower and
less frequent dosing of patients. Thus, enhanced stability in human
liver microsomes is a favorable characteristic for compounds that
are to be used for drugs. Therefore, compounds of the present
invention in addition to being able to inhibit TRPA1 are expected
to have a favorable in vivo clearance and thus the desired duration
of action in humans.
Stability in human hepatocytes refers to the susceptibility of
compounds to biotransformation in the context of selecting and/or
designing drugs with favorable pharmacokinetic properties. The
primary site of metabolism for many drugs is the liver. Human
hepatocytes contain the cytochrome P450s (CYPs) and other drug
metabolizing enzymes, and thus represent a model system for
studying drug metabolism in vitro. (Importantly, in contrast to
liver microsomes assay, the hepatocytes assay covers also phase II
biotransformations as well as liver-specific transporter-mediated
processes, and therefore represents a more complete system for drug
metabolism studies.) Enhanced stability in human hepatocytes is
associated with several advantages, including increased
bioavailability and adequate half-life, which can enable lower and
less frequent dosing of patients. Thus, enhanced stability in human
hepatocytes is a favorable characteristic for compounds that are to
be used for drugs.
The present invention provides novel compounds according to formula
(I)
##STR00003## wherein
A is selected from the group consisting of phenyl, naphthyl,
thiophenyl, benzothiophenyl or benzofuranyl, optionally substituted
with one or two members of the group consisting of H, F, Cl, Br,
C.sub.1-4-alkyl, F.sub.1_3-fluoro-C.sub.1-4-alkyl, CN, OCH.sub.3,
cyclopropyl, and cyclobutyl,
or
A is selected from
##STR00004## and
R.sup.1 is selected from H, C.sub.1-4-alkyl,
F.sub.1-3-fluoro-C.sub.1-4-alkyl, C.sub.1-4-alkyl-OH or
C.sub.1-4-alkyl-CN;
R.sup.2 is selected from C.sub.1-2-alkyl or Cl;
or R.sup.1 and R.sup.2 are each CH.sub.2 joined via a bond forming
a 6-membered ring.
Another embodiment of the present invention relates to a compound
of formula (I), wherein A is selected from the group consisting of
phenyl or benzofuranyl, optionally substituted with one or two
members of the group consisting of H, F, Cl, Br, C.sub.1-4-alkyl,
F.sub.1-3-fluoro-C.sub.1-4-alkyl, CN, OCH.sub.3, cyclopropyl, and
cyclobutyl,
or
A is selected from
##STR00005## and substituents R.sup.1 and R.sup.2 are defined as in
any of the preceding embodiments.
Another embodiment of the present invention relates to a compound
of formula (I), wherein A is selected from the group consisting of
phenyl or benzofuranyl, optionally substituted with one or two
members of the group consisting of H, F, Cl, and CH.sub.3;
or
A is selected from
##STR00006## and substituents R.sup.1 and R.sup.2 are defined as in
any of the preceding embodiments.
Another embodiment of the present invention relates to a compound
of formula (I), wherein A is selected from the group consisting
of
##STR00007## and substituents R.sup.1 and R.sup.2 are defined as in
any of the preceding embodiments.
Another embodiment of the present invention relates to a compound
of formula (I), wherein R.sup.1 is selected from the group
consisting of H, CH.sub.3, CH.sub.2CHF.sub.2 or
CH.sub.2C(CH.sub.3).sub.2OH; and substituents A and R.sup.2 are
defined as in any of the preceding embodiments.
Another embodiment of the present invention relates to a compound
of formula (I), wherein R.sup.2 is --CH.sub.3; and substituents A
and R.sup.1 are defined as in any of the preceding embodiments.
Preferred is a compound of formula (I), selected from the group
consisting of
##STR00008## and substituent A is defined as in any of the
preceding embodiments.
Particularly preferred is the (S)-enantiomer of a compound
according to formula (I) selected from the group consisting of
##STR00009## ##STR00010##
USED TERMS AND DEFINITIONS
Terms not specifically defined herein should be given the meanings
that would be given to them by one of skill in the art in light of
the disclosure and the context. As used in the specification,
however, unless specified to the contrary, the following terms have
the meaning indicated and the following conventions are adhered
to.
In the groups, radicals, or moieties defined below, the number of
carbon atoms is often specified preceding the group, for example,
C.sub.1-6-alkyl means an alkyl group or radical having 1 to 6
carbon atoms. In general in groups like HO, H.sub.2N, (O)S,
(O).sub.2S, NC (cyano), HOOC, F.sub.3C or the like, the skilled
artisan can see the radical attachment point(s) to the molecule
from the free valences of the group itself. For combined groups
comprising two or more subgroups, the last named subgroup is the
radical attachment point, for example, the substituent
"aryl-C.sub.1-3-alkyl" means an aryl group which is bound to a
C.sub.1-3-alkyl-group, the latter of which is bound to the core or
to the group to which the substituent is attached.
In case a compound of the present invention is depicted in form of
a chemical name and as a formula in case of any discrepancy the
formula shall prevail. An asterisk is may be used in sub-formulas
to indicate the bond which is connected to the core molecule as
defined.
The numeration of the atoms of a substituent starts with the atom
that is closest to the core or to the group to which the
substituent is attached.
For example, the term "3-carboxypropyl-group" represents the
following substituent:
##STR00011## wherein the carboxy group is attached to the third
carbon atom of the propyl group. The terms "1-methylpropyl-",
"2,2-dimethylpropyl-" or "cyclopropylmethyl-" group represent the
following groups:
##STR00012##
The asterisk may be used in sub-formulas to indicate the bond that
is connected to the core molecule as defined.
The term "C.sub.1-n-alkyl", wherein n is an integer selected from
2, 3, 4 or 5, either alone or in combination with another radical
denotes an acyclic, saturated, branched or linear hydrocarbon
radical with 1 to n C atoms. For example the term C.sub.1-5-alkyl
embraces the radicals H.sub.3C--, H.sub.3C--CH.sub.2--,
H.sub.3C--CH.sub.2--CH.sub.2--, H.sub.3C--CH(CH.sub.3)--,
H.sub.3C--CH.sub.2--CH.sub.2--CH.sub.2--,
H.sub.3C--CH.sub.2--CH(CH.sub.3)--,
H.sub.3C--CH(CH.sub.3)--CH.sub.2--, H.sub.3C--C(CH.sub.3).sub.2--,
H.sub.3C--CH.sub.2--CH.sub.2--CH.sub.2--CH.sub.2--,
H.sub.3C--CH.sub.2--CH.sub.2--CH(CH.sub.3)--,
H.sub.3C--CH.sub.2--CH(CH.sub.3)--CH.sub.2--,
H.sub.3C--CH(CH.sub.3)--CH.sub.2--CH.sub.2--,
H.sub.3C--CH.sub.2--C(CH.sub.3).sub.2--,
H.sub.3C--C(CH.sub.3).sub.2--CH.sub.2--,
H.sub.3C--CH(CH.sub.3)--CH(CH.sub.3)-- and
H.sub.3C--CH.sub.2--CH(CH.sub.2CH.sub.3)--.
The term "F.sub.1-m-fluoro-C.sub.1-n-alkyl", wherein m is an
integer selected from 2 or 3, and n is an integer selected from 2,
3, 4 or 5, denotes an C.sub.1-n-alkyl group as defined hereinbefore
wherein one or more hydrogen atoms are replaced by 1-m fluorine.
For example, F.sub.1-3-fluoro-C.sub.1-2-alkyl embraces the radicals
FH.sub.2C--, F.sub.2HC--, F.sub.3C--, FH.sub.2C--H.sub.2C--,
F.sub.2HC--H.sub.2C--, F.sub.3C--H.sub.2C--, FH.sub.2C--FHC--,
FH.sub.2C--F.sub.2C--, F.sub.2HC--FHC--, H.sub.3C--FHC--, and
H.sub.3C--F.sub.2C--.
The term phenyl refers to the radical of the following ring
##STR00013##
The term naphthyl refers to the radical of the following ring
##STR00014##
The term thiophenyl refers to the radical of the following ring
##STR00015##
The term benzothiophenyl refers to the radical of the following
ring
##STR00016##
The term thienopyrimidone refers to the radical of the following
ring
##STR00017## and includes
##STR00018##
The term benzofuranyl refers to the radical of the following
ring
##STR00019##
The term cyclopropyl refers to the radical of the following
ring
##STR00020##
The term cyclobutyl refers to the radical of the following ring
##STR00021##
The term "substituted" as used herein, means that any one or more
hydrogens on the designated atom is replaced with a selection from
the indicated group, provided that the designated atom's normal
valence is not exceeded, and that the substitution results in a
stable compound.
Unless specifically indicated, throughout the specification and the
appended claims, a given chemical formula or name shall encompass
tautomers and all stereo, optical and geometrical isomers (e.g.
enantiomers, diastereomers, E/Z isomers etc.) and racemates thereof
as well as mixtures in different proportions of the separate
enantiomers, mixtures of diastereomers, or mixtures of any of the
foregoing forms where such isomers and enantiomers exist, as well
as salts, including pharmaceutically acceptable salts thereof and
solvates thereof such as for instance hydrates including solvates
of the free compounds or solvates of a salt of the compound.
In general, substantially pure stereoisomers can be obtained
according to synthetic principles known to a person skilled in the
field, e.g. by separation of corresponding mixtures, by using
stereochemically pure starting materials and/or by stereoselective
synthesis. It is known in the art how to prepare optically active
forms, such as by resolution of racemic forms or by synthesis, e.g.
starting from optically active starting materials and/or by using
chiral reagents.
Enantiomerically pure compounds of this invention or intermediates
may be prepared via asymmetric synthesis, for example by
preparation and subsequent separation of appropriate diastereomeric
compounds or intermediates which can be separated by known methods
(e.g. by chromatographic separation or crystallization) and/or by
using chiral reagents, such as chiral starting materials, chiral
catalysts or chiral auxiliaries.
Further, it is known to the person skilled in the art how to
prepare enantiomerically pure compounds from the corresponding
racemic mixtures, such as by chromatographic separation of the
corresponding racemic mixtures on chiral stationary phases; or by
resolution of a racemic mixture using an appropriate resolving
agent, e.g. by means of diastereomeric salt formation of the
racemic compound with optically active acids or bases, subsequent
resolution of the salts and release of the desired compound from
the salt; or by derivatization of the corresponding racemic
compounds with optically active chiral auxiliary reagents,
subsequent diastereomer separation and removal of the chiral
auxiliary group; or by kinetic resolution of a racemate (e.g. by
enzymatic resolution); by enantioselective crystallization from a
conglomerate of enantiomorphous crystals under suitable conditions;
or by (fractional) crystallization from a suitable solvent in the
presence of an optically active chiral auxiliary.
The phrase "pharmaceutically acceptable" is employed herein to
refer to those compounds, materials, compositions, and/or dosage
forms which are, within the scope of sound medical judgment,
suitable for use without excessive toxicity, irritation, allergic
response, or other problem or complication, and commensurate with a
reasonable benefit/risk ratio.
As used herein, "pharmaceutically acceptable salt" refers to
derivatives of the disclosed compounds wherein the parent compound
forms a salt or a complex with an acid or a base. Examples of acids
forming a pharmaceutically acceptable salt with a parent compound
containing a basic moiety include mineral or organic acids such as
benzenesulfonic acid, benzoic acid, citric acid, ethanesulfonic
acid, fumaric acid, gentisic acid, hydrobromic acid, hydrochloric
acid, maleic acid, malic acid, malonic acid, mandelic acid,
methanesulfonic acid, 4-methyl-benzenesulfonic acid, phosphoric
acid, salicylic acid, succinic acid, sulfuric acid and tartaric
acid.
Examples for cations and bases forming a pharmaceutically
acceptable salt with a parent compound containing an acidic moiety
include Na.sup.+, K.sup.+, Ca.sup.2+, Mg.sup.2+, NH.sub.4.sup.+,
L-arginine, 2,2'-iminobisethanol, L-lysine, N-methyl-D-glucamine or
tris(hydroxymethyl)-aminomethane. The pharmaceutically acceptable
salts of the present invention can be synthesized from the parent
compound that contains a basic or acidic moiety by conventional
chemical methods. Generally, such salts can be prepared by reacting
the free acid or base forms of these compounds with a sufficient
amount of the appropriate base or acid in water or in an organic
diluent like ether, ethyl acetate, ethanol, isopropanol, or
acetonitrile, or a mixture thereof.
Salts of other acids than those mentioned above which for example
are useful for purifying or isolating the compounds of the present
invention (e.g. trifluoroacetate salts,) also comprise a part of
the present invention.
BIOLOGICAL ASSAYS
Assay A: TRPA1 Assay
The activity of the compounds of the invention may be demonstrated
using the following in vitro TRPA1 cell assay:
Method:
A human HEK293 cell line over-expressing the human TRPA1 ion
channel (Perkin Elmer, Product No. AX-004-PCL) was used as a test
system for compound efficacy and potency. Compound activity was
determined by measuring the effect of compounds on intracellular
calcium concentration induced by AITC (Allylisothiocyanat) agonism
in a FLIPRtetra system (Molecular Devices).
Cell Culture:
The cells were obtained as frozen cells in cryo-vials and stored
until use at -150.degree. C. Cells were grown in culture medium
(MEM/EBSS medium with 10% FCS and 0.4 mg/ML Geneticin). It is
important that density does not exceed 90% confluence. For
sub-culturing cells were detached from flasks by Versene. At the
day before the assay, cells were detached, washed twice with medium
(MEM/EBSS medium with 10% FCS) and 20000 cells in 20p/well were
seeded to Poly D-Lysin biocoated 384-well plates (black, clear
bottom, Cat. 356697) from Corning. Plates were incubated for 24
hours at 37.degree. C./5% CO2 before use in the assay.
Compound Preparation
The test compounds were dissolved in 100% DMSO at a concentration
of 10 mM and in a first step diluted in DMSO to a concentration of
5 mM, followed by serial dilution steps in 100% DMSO. Dilution
factor and number of dilution steps may vary according to needs.
Typically 8 different concentrations by 1:5 dilutions were
prepared, further intermediate dilutions (1:20) of the substances
were carried out with HBSS/HEPES buffer (1.times.HEPES, Cat.14065
from Gibco, 20 mM HEPES, Cat. 83264 from SIGMA, 0.1% BSA Cat.11926
from Invitrogen, pH 7,4
FLIPR Assay:
At the assay day cells were washed 3.times. with assay puffer, 20
.mu.L buffer remained in the wells after washing. 10 .mu.L Ca6 kit
(Cat.R8191 MolecularDevices) loading buffer in HBSS/HEPES was added
to the cells and the plates were incubated with lid for 120 minutes
at 37.degree./5% CO2. 10 .mu.L of compound or controls in
HBSS/HEPES buffer/5% DMSO from the intermediate dilution plate were
carefully added to the wells. Luminescence (indicating the calcium
influx or release) was read on the FLIPRtetra device for 10 minutes
to monitor the compound induced effects (e.g. agonism). Finally 10
.mu.L of the agonist AITC 50 .mu.M dissolved in HBSS/HEPES
buffer/0.05% DMSO (final concentration 10 .mu.M) was added to the
wells followed by an additional read on the FLIPRtetra device for
10 minutes. The area under the signal curve (AUC) after AITC
addition was used for IC.sub.50/% inhibition calculations
Data Evaluation and Calculation:
Each assay microtiter plate contained wells with vehicle (1% DMSO)
controls instead of compound as controls for AITC induced
luminescence (100% CTL; high controls) and wells with vehicle
controls without AITC as controls for non-specific changes in
luminescence (0% CTL; low controls).
The analysis of the data was performed by the calculation of the
area under signal curve of the individual wells. Based on this
values the % value for the measurement of each substance
concentration was calculated
(AUC(sample)-AUC(low))*100/(AUC(high)-AUC(low)) using MegaLab
software (in house development). The IC50 values were calculated
from the % control values using MegaLab software. Calculation:
[y=(ad)/(1+(x/c){circumflex over ( )}b)+d], a=low value, d=high
value; x=conc M; c=IC50 M; b=hill; y=% ctrl
TABLE-US-00001 TABLE 1 Biological data for compounds of the
invention as obtained in Assay A Example hTRPA1 IC.sub.50 [nM] 1 13
2 20 3 19 4 33 5 48 6 48 7 34 8 56 9 78 10 52 11 10 12 15 13 37 14
33 15 9 16 16
TABLE-US-00002 TABLE 2 Biological data for prior art compounds
(examples 53, 72, 73, 86, 90 in WO2017/060488) as obtained in Assay
A. Example in WO2017/060488 hTRPA1 IC.sub.50 [nM] 53 36 72 14 73 35
86 67 90 41
TABLE-US-00003 TABLE 3 Biological data for prior art compounds
(example 31 in L. Schenkel, et al., J. Med. Chem. 2016, 59,
2794-2809) as obtained in Assay A. Example in Med. Chem. 2016, 59,
2794-2809 hTRPA1 IC.sub.50 [nM] 31 52
Assay B: Microsomal Clearance:
The metabolic degradation of the test compound is assayed at
37.degree. C. with pooled liver microsomes. The final incubation
volume of 100 .mu.l per time point contains TRIS buffer pH 7.6 at
RT (0.1 M), magnesium chloride (5 mM), microsomal protein (1 mg/ml)
and the test compound at a final concentration of 1 .mu.M.
Following a short preincubation period at 37.degree. C., the
reactions are initiated by addition of beta-nicotinamide adenine
dinucleotide phosphate, reduced form (NADPH, 1 mM) and terminated
by transferring an aliquot into solvent after different time points
(0, 5, 15, 30, 60 min). Additionally, the NADPH-independent
degradation is monitored in incubations without NADPH, terminated
at the last time point. The [%] remaining test compound after NADPH
independent incubation is reflected by the parameter c(control)
(metabolic stability). The quenched incubations are pelleted by
centrifugation (10000 g, 5 min).
An aliquot of the supernatant is assayed by LC-MS/MS for the amount
of parent compound. The half-life (t1/2 INVITRO) is determined by
the slope of the semilogarithmic plot of the concentration-time
profile.
The intrinsic clearance (CL_INTRINSIC) is calculated by considering
the amount of protein in the incubation: CL_INTRINSIC [.mu.l/min/mg
protein]=(Ln 2/(half-life [min]*protein content [mg/ml]))*1000
CL_INTRINSIC_INVIVO [ml/min/kg]=(CL_INTRINSIC [.mu.L/min/mg
protein].times.MPPGL [mg protein/g liver].times.liver factor [g/kg
bodyweight])/1000 Qh [%]=CL [ml/min/kg]/hepatic blood flow
[ml/min/kg]) Hepatocellularity, human: 120.times.10e6 cells/g liver
Liver factor, human: 25.7 g/kg bodyweight Blood flow, human: 21
ml/(min.times.kg)
TABLE-US-00004 TABLE 4 Biological data for compounds of the
invention as obtained in Assay B Example human LM [% Qh] 1 29 2
<23 3 <23 4 <23 5 <23 6 <23 7 23 8 <23 9 29 10
<23 11 <23 12 24 13 <23 14 <23 15 <23 16 <23
TABLE-US-00005 TABLE 5 Biological data for prior art compounds
(examples 53, 72, 73, 86, 90 in WO2017/060488) as obtained in Assay
B. Example in WO2017/060488 human LM [% Qh] 53 <23 72 30 73 38
86 <23 90 39
TABLE-US-00006 TABLE 6 Biological data for prior art compounds
(example 31 in L. Schenkel, et al., J. Med. Chem. 2016, 59,
2794-2809) as obtained in Assay B. Example in Med. Chem. 2016, 59,
2794-2809 human LM [% Qh] 31 <23
Assay C: Hepatocyte Clearance
The metabolic degradation of the test compound is assayed in a
hepatocyte suspension. Hepatocytes (cryopreserved) are incubated
in. Dulbecco's modified eagle medium (supplemented with 3.5 .mu.g
glucagon/500 mL, 2.5 mg insulin/500 mL and 3.75 mg/500 mL
hydrocortison) containing 5% or 50% species serum.
Following a 30 min preincubation in an incubator (37.degree. C.,
10% CO2) 5 .mu.l of test compound solution (80 .mu.M; from 2 mM in
DMSO stock solution diluted 1:25 with medium) are added into 395
.mu.l hepatocyte suspension (cell density in the range 0.25-5 Mio
cells/mL depending on the species, typically 1 Mio cells/mL; final
concentration of test compound 1 .mu.M, final DMSO concentration
0.05%).
The cells are incubated for six hours (incubator, orbital shaker)
and samples (25 .mu.l) are taken at 0, 0.5, 1, 2, 4 and 6 hours.
Samples are transferred into acetonitrile and pelleted by
centrifugation (5 min). The supernatant is transferred to a new
96-deepwell plate, evaporated under nitrogen and resuspended.
Decline of parent compound is analyzed by HPLC-MS/MS
CLint is calculated as follows
CL_INTRINSIC=Dose/AUC=(C0/CD)/(AUD+clast/k).times.1000/60. C0:
initial concentration in the incubation [.mu.M], CD: cell density
of vital cells [10e6cells/mL], AUD: area under the data
[.mu.M.times.h], clast: concentration of last data point [.mu.M],
k: slope of the regression line for parent decline [h-1].
The calculated in vitro hepatic intrinsic clearance can be scaled
up to the intrinsic in vivo hepatic Clearance and used to predict
hepatic in vivo blood clearance (CL) by the use of a liver model
(well stirred model). CL_INTRINSIC_INVIVO [ml/min/kg]=(CL_INTRINSIC
[.mu.L/min/10e6cells].times.hepatocellularity [10e6 cells/g
liver].times.liver factor [g/kg bodyweight])/1000 CL
[ml/min/kg]=CL_INTRINSIC_INVIVO [ml/min/kg].times.hepatic blood
flow [ml/min/kg]/(CL_INTRINSIC_INVIVO [ml/min/kg]+hepatic blood
flow [ml/min/kg]) Qh [%]=CL [ml/min/kg]/hepatic blood flow
[ml/min/kg]) Hepatocellularity, human: 120.times.10e6 cells/g liver
Liver factor, human: 25.7 g/kg bodyweight Blood flow, human: 21
ml/(min.times.kg)
TABLE-US-00007 TABLE 7 Biological data for compounds of the
invention as obtained in Assay C human Hepatocytes Example [% Qh] 1
25 2 29 3 36 4 29 5 37 6 23 7 39 8 24 9 24 10 25 11 29 12 36 13 18
14 24 15 31 16 39
TABLE-US-00008 TABLE 8 Biological data for prior art compounds
(examples 53, 72, 73, 86, 90 in WO2017/060488) as obtained in Assay
C. Example in human Hepatocytes WO2017/060488 [% Qh] 53 25 72 50 73
36 86 12 90 61
TABLE-US-00009 TABLE 9 Biological data for prior art compounds
(example 31 in L. Schenkel, et al., J. Med. Chem. 2016, 59,
2794-2809) as obtained in Assay C. Example in Med. Chem. human
Hepatocytes 2016, 59, 2794-2809 [% Qh] 31 73
METHOD OF TREATMENT
The present invention is directed to compounds of general formula 1
which are useful in the prevention and/or treatment of a disease
and/or condition associated with or modulated by TRPA1 activity,
including but not limited to the treatment and/or prevention of
fibrotic disease, inflammatory and immunoregulatory disorders,
respiratory or gastrointestinal diseases or complaints, ophthalmic
diseases, inflammatory diseases of the joints and inflammatory
diseases of the nasopharynx, eyes, and skin. Said disorders,
diseases and complaints include cough, idiopathic pulmonary
fibrosis, other pulmonary interstitial diseases and other fibrotic,
asthma or allergic diseases, eosinophilic diseases, chronic
obstructive pulmonary disease, as well as autoimmune pathologies,
such as rheumatoid arthritis and atherosclerosis, pain and
neurological disorders, such as depression.
The compounds of general formula 1 are useful for the prevention
and/or treatment of:
(1) Cough such as chronic idiopathic cough or chronic refractory
cough, cough associated with asthma, COPD and lung cancer and
post-viral cough.
(2) Pulmonary fibrotic diseases such as pneumonitis or interstitial
pneumonitis associated with collagenosis, e.g. lupus erythematodes,
systemic scleroderma, rheumatoid arthritis, polymyositis and
dermatomysitis, idiopathic interstitial pneumonias, such as
pulmonary lung fibrosis (IPF), non-specific interstitial pneumonia,
respiratory bronchiolitis associated interstitial lung disease,
desquamative interstitial pneumonia, cryptogenic organizing
pneumonia, acute interstitial pneumonia and lymphocytic
interstitial pneumonia, lymangioleiomyomatosis, pulmonary alveolar
proteinosis, Langerhan's cell histiocytosis, pleural parenchymal
fibroelastosis, interstitial lung diseases of known cause, such as
interstitial pneumonitis as a result of occupational exposures such
as asbestosis, silicosis, miners lung (coal dust), farmers lung
(hay and mould), Pidgeon fanciers lung (birds) or other
occupational airbourne triggers such as metal dust or mycobacteria,
or as a result of treatment such as radiation, methotrexate,
amiodarone, nitrofurantoin or chemotherapeutics, or for
granulomatous disease, such as granulomatosis with polyangitis,
Churg-Strauss syndrome, sarcoidosis, hypersensitivity pneumonitis,
or interstitial pneumonitis caused by different origins, e.g.
aspiration, inhalation of toxic gases, vapors, bronchitis or
pneumonitis or interstitial pneumonitis caused by heart failure,
X-rays, radiation, chemotherapy, M. boeck or sarcoidosis,
granulomatosis, cystic fibrosis or mucoviscidosis, or
alpha-1-antitrypsin deficiency. (3) Other fibrotic diseases such as
hepatic bridging fibrosis, liver cirrhosis, non-alcoholic
steatohepatitis (NASH), atrial fibrosis, endomyocardial fibrosis,
old myocardial infarction, glial scar, arterial stiffness,
arthrofibrosis, Dupuytren's contracture, keloid,
scleroderma/systemic sclerosis, mediastinal fibrosis,
myelofibrosis, Peyronie's disease, nephrogenic systemic fibrosis,
retroperitoneal fibrosis, adhesive capsulitis. (4) Inflammatory,
auto-immune or allergic diseases and conditions such as allergic or
non-allergic rhinitis or sinusitis, chronic sinusitis or rhinitis,
nasal polyposis, chronic rhinosinusitis, acute rhinosinusitis,
asthma, pediatric asthma, allergic bronchitis, alveolitis,
hyperreactive airways, allergic conjunctivitis, bronchiectasis,
adult respiratory distress syndrome, bronchial and pulmonary edema,
bronchitis or pneumonitis, eosinophilic cellulites (e.g., Well's
syndrome), eosinophilic pneumonias (e.g., Loeffler's syndrome,
chronic eosinophilic pneumonia), eosinophilic fasciitis (e. g.,
Shulman's syndrome), delayed-type hypersensitivity, non-allergic
asthma; exercise induced bronchoconstriction; chronic obstructive
pulmonary disease (COPD), acute bronchitis, chronic bronchitis,
cough, pulmonary emphysema; systemic anaphylaxis or
hypersensitivity responses, drug allergies (e.g., to penicillin,
cephalosporin), eosinophiliamyalgia syndrome due to the ingestion
of contaminated tryptophane, insect sting allergies; autoimmune
diseases, such as rheumatoid arthritis, Graves' disease, Sjogren's
syndrome psoriatic arthritis, multiple sclerosis, systemic lupus
erythematosus, myasthenia gravis, immune thrombocytopenia (adult
ITP, neonatal thrombocytopenia, pediatric ITP), immune hemolytic
anemia (auto-immune and drug induced), Evans syndrome (platelet and
red cell immune cytopaenias), Rh disease of the newborn,
Goodpasture's syndrome (anti-GBM disease), Celiac, autoimmune
cardio-myopathy juvenile onset diabetes; glomerulonephritis,
autoimmune thyroiditis, Behcet's disease; graft rejection (e.g., in
transplantation), including allograft rejection or graftversus-host
disease; inflammatory bowel diseases, such as Crohn's disease and
ulcerative colitis; spondyloarthropathies; scleroderma; psoriasis
(including T-cell mediated psoriasis) and inflammatory dermatoses
such as an dermatitis, eczema, atopic dermatitis, allergic contact
dermatitis, urticaria; vasculitis (e. g., necrotizing, cutaneous,
and hypersensitivity vasculitis); erythema nodosum; eosinophilic
myositis, eosinophilic fasciitis, cancers with leukocyte
infiltration of the skin or organs; ophthalmic diseases such as age
related macular degeneration, diabetic retinopathy and diabetic
macular edema, keratitis, eosinophilic keratitis,
keratoconjunctivitis, vernal keratoconjunctivitis, scarring,
anterior segment scarring, blepharitis, blepharoconjunctivitis,
bullous disorders, cicatricial pemphigoid, conjunctival melanoma,
papillary conjunctivitis, dry eye, episcleritis, glaucoma, gliosis,
Granuloma annulare, Graves' ophthalmopathy, intraocular melanoma,
Pinguecula, proliferative vitreoretinopathy, pterygia, scleritis,
uveitis, acute gout flares, gout or osteoarthritis. (5) Pain such
as chronic idiopathic pain syndrome, neuropathic pain, dysesthesia,
allodynia, migraine, dental pain and post-surgical pain. (6)
Depression, anxiousness, diabetic neuropathy and bladder disorders
such as bladder outlet obstruction, overactive bladder, cystitis;
myocardial reperfusion injury or brain ischaemia injury.
Accordingly, the present invention relates to a compound of general
formula 1 for use as a medicament.
Furthermore, the present invention relates to the use of a compound
of general formula 1 for the treatment and/or prevention of a
disease and/or condition associated with or modulated by TRPA1
activity.
Furthermore, the present invention relates to the use of a compound
of general formula 1 for the treatment and/or prevention of
fibrotic disease, inflammatory and immunoregulatory disorders,
respiratory or gastrointestinal diseases or complaints, ophthalmic
diseases, inflammatory diseases of the joints and inflammatory
diseases of the nasopharynx, eyes, and skin. Said disorders,
diseases and complaints include cough, idiopathic pulmonary
fibrosis, other pulmonary interstitial diseases and other fibrotic,
asthma or allergic diseases, eosinophilic diseases, chronic
obstructive pulmonary disease, as well as autoimmune pathologies,
such as rheumatoid arthritis and atherosclerosis.
Furthermore, the present invention relates to the use of a compound
of general formula 1 for the treatment and/or prevention of:
(1) Cough such as chronic idiopathic cough or chronic refractory
cough, cough associated with asthma, COPD and lung cancer and
post-viral cough.
(2) Pulmonary fibrotic diseases such as pneumonitis or interstitial
pneumonitis associated with collagenosis, e.g. lupus erythematodes,
systemic scleroderma, rheumatoid arthritis, polymyositis and
dermatomysitis, idiopathic interstitial pneumonias, such as
pulmonary lung fibrosis (IPF), non-specific interstitial pneumonia,
respiratory bronchiolitis associated interstitial lung disease,
desquamative interstitial pneumonia, cryptogenic organizing
pneumonia, acute interstitial pneumonia and lymphocytic
interstitial pneumonia, lymangioleiomyomatosis, pulmonary alveolar
proteinosis, Langerhan's cell histiocytosis, pleural parenchymal
fibroelastosis, interstitial lung diseases of known cause, such as
interstitial pneumonitis as a result of occupational exposures such
as asbestosis, silicosis, miners lung (coal dust), farmers lung
(hay and mould), Pidgeon fanciers lung (birds) or other
occupational airbourne triggers such as metal dust or mycobacteria,
or as a result of treatment such as radiation, methotrexate,
amiodarone, nitrofurantoin or chemotherapeutics, or for
granulomatous disease, such as granulomatosis with polyangitis,
Churg-Strauss syndrome, sarcoidosis, hypersensitivity pneumonitis,
or interstitial pneumonitis caused by different origins, e.g.
aspiration, inhalation of toxic gases, vapors, bronchitis or
pneumonitis or interstitial pneumonitis caused by heart failure,
X-rays, radiation, chemotherapy, M. boeck or sarcoidosis,
granulomatosis, cystic fibrosis or mucoviscidosis, or
alpha-1-antitrypsin deficiency. (3) Other fibrotic diseases such as
heaptic bridging fibrosis, liver cirrhosis, non-alcoholic
steatohepatitis (NASH), atrial fibrosis, endomyocardial fibrosis,
old myocardial infarction, glial scar, arterial stiffness,
arthrofibrosis, Dupuytren's contracture, keloid,
scleroderma/systemic sclerosis, mediastinal fibrosis,
myelofibrosis, Peyronie's disease, nephrogenic systemic fibrosis,
retroperitoneal fibrosis, adhesive capsulitis. (4) Inflammatory,
auto-immune or allergic diseases and conditions such as allergic or
non-allergic rhinitis or sinusitis, chronic sinusitis or rhinitis,
nasal polyposis, chronic rhinosinusitis, acute rhinosinusitis,
asthma, pediatric asthma, allergic bronchitis, alveolitis,
hyperreactive airways, allergic conjunctivitis, bronchiectasis,
adult respiratory distress syndrome, bronchial and pulmonary edema,
bronchitis or pneumonitis, eosinophilic cellulites (e.g., Well's
syndrome), eosinophilic pneumonias (e.g., Loeffler's syndrome,
chronic eosinophilic pneumonia), eosinophilic fasciitis (e. g.,
Shulman's syndrome), delayed-type hypersensitivity, non-allergic
asthma; exercise induced bronchoconstriction; chronic obstructive
pulmonary disease (COPD), acute bronchitis, chronic bronchitis,
cough, pulmonary emphysema; systemic anaphylaxis or
hypersensitivity responses, drug allergies (e.g., to penicillin,
cephalosporin), eosinophiliamyalgia syndrome due to the ingestion
of contaminated tryptophane, insect sting allergies; autoimmune
diseases, such as rheumatoid arthritis, Graves' disease, Sjogren's
syndrome psoriatic arthritis, multiple sclerosis, systemic lupus
erythematosus, myasthenia gravis, immune thrombocytopenia (adult
ITP, neonatal thrombocytopenia, pediatric ITP), immune hemolytic
anemia (auto-immune and drug induced), Evans syndrome (platelet and
red cell immune cytopaenias), Rh disease of the newborn,
Goodpasture's syndrome (anti-GBM disease), Celiac, autoimmune
cardio-myopathy juvenile onset diabetes; glomerulonephritis,
autoimmune thyroiditis, Behcet's disease; graft rejection (e.g., in
transplantation), including allograft rejection or graftversus-host
disease; inflammatory bowel diseases, such as Crohn's disease and
ulcerative colitis; spondyloarthropathies; scleroderma; psoriasis
(including T-cell mediated psoriasis) and inflammatory dermatoses
such as an dermatitis, eczema, atopic dermatitis, allergic contact
dermatitis, urticaria; vasculitis (e. g., necrotizing, cutaneous,
and hypersensitivity vasculitis); erythema nodosum; eosinophilic
myositis, eosinophilic fasciitis, cancers with leukocyte
infiltration of the skin or organs; ophthalmic diseases such as age
related macular degeneration, diabetic retinopathy and diabetic
macular edema, keratitis, eosinophilic keratitis,
keratoconjunctivitis, vernal keratoconjunctivitis, scarring,
anterior segment scarring, blepharitis, blepharoconjunctivitis,
bullous disorders, cicatricial pemphigoid, conjunctival melanoma,
papillary conjunctivitis, dry eye, episcleritis, glaucoma, gliosis,
Granuloma annulare, Graves' ophthalmopathy, intraocular melanoma,
Pinguecula, proliferative vitreoretinopathy, pterygia, scleritis,
uveitis, acute gout flares, gout or osteoarthritis. (5) Pain such
as chronic idiopathic pain syndrome, neuropathic pain, dysesthesia,
allodynia, migraine, dental pain and post-surgical pain. (6)
Depression, anxiousness, diabetic neuropathy and bladder disorders
such as bladder outlet obstruction, overactive bladder, cystitis;
myocardial reperfusion injury or brain ischaemia injury.
In a further aspect the present invention relates to a compound of
general formula 1 for use in the treatment and/or prevention of
above mentioned diseases and conditions.
In a further aspect the present invention relates to the use of a
compound of general formula 1 for the preparation of a medicament
for the treatment and/or prevention of above mentioned diseases and
conditions.
In a further aspect of the present invention the present invention
relates to methods for the treatment or prevention of above
mentioned diseases and conditions, which method comprises the
administration of an effective amount of a compound of general
formula 1 to a human being.
COMBINATION THERAPY
The compounds of the invention may further be combined with one or
more, preferably one additional therapeutic agent. According to one
embodiment the additional therapeutic agent is selected from the
group of therapeutic agents useful in the treatment of diseases or
conditions described hereinbefore, in particular associated with
fibrotic diseases, inflammatory and immunoregulatory disorders,
respiratory or gastrointestinal diseases or complaints,
inflammatory diseases of the joints or of the nasopharynx, eyes,
and skin or conditions such as for example cough, idiopathic
pulmonary fibrosis, other pulmonary interstitial diseases, asthma
or allergic diseases, eosinophilic diseases, chronic obstructive
pulmonary disease, atopic dermatitis as well as autoimmune
pathologies, such as rheumatoid arthritis and atherosclerosis, or
therapeutic agents useful for the treatment of ophthalmic diseases,
pain and depression.
Additional therapeutic agents that are suitable for such
combinations include in particular those, which, for example,
potentiate the therapeutic effect of one or more active substances
with respect to one of the indications mentioned and/or allow the
dosage of one or more active substances to be reduced.
Therefore, a compound of the invention may be combined with one or
more additional therapeutic agents selected from the group
consisting of antifibrotic agents, anti-tussive agents,
anti-inflammatory agents, anti-atopic dermatitis agents,
analgesics, anti-convulsants, anxiolytics, sedatives, skeletal
muscle relaxants or anti-depressants.
Antifibrotic agents are for example nintedanib, pirfenidone,
phosphodiesterase-IV (PDE4) inhibitors such as roflumilast,
autotaxin inhibitors such as GLPG-1690 or BBT-877; connective
tissue growth factor (CTGF) blocking antibodies such as
Pamrevlumab; B-cell activating factor receptor (BAFF-R) blocking
antibodies such as Lanalumab; alpha-V/beta-6 blocking inhibitors
such as BG-00011/STX-100, recombinant pentraxin-2 (PTX-2) such as
PRM-151; c-Jun N-terminal kinase (JNK) inhibitors such as CC-90001;
galectin-3 inhibitors such as TD-139; G-protein coupled receptor 84
(GPR84) inhibitors such as GLPG1205; G-protein coupled receptor
84/G-protein coupled receptor 40 dual inhibitors such as PBI-4050;
Rho Associated Coiled-Coil Containing Protein Kinase 2 (ROCK2)
inhibitors such as KD-025; heat shock protein 47 (HSP47) small
interfering RNA such as BMS986263/ND-L02-s0201; Wnt pathway
inhibitor such as SM-04646; LD4/PDE3/4 inhibitors such as
Tipelukast; recombinant immuno-modulatory domains of histidyl tRNA
synthetase (HARS) such as ATYR-1923; prostaglandin synthase
inhibitors such as ZL-2102/SAR-191801; 15-hydroxy-eicosapentaenoic
acid (15-HEPE e.g. DS-102); Lysyl Oxidase Like 2 (LOXL2) inhibitors
such as PAT-1251, PXS-5382/PXS-5338; phosphoinositide 3-kinases
(PI3K)/mammalian target of rapamycin (mTOR) dual inhibitors such as
HEC68498; calpain inhibitors such as BLD-2660; mitogen-activated
protein kinase kinase kinase (MAP3K19) inhibitors such as
MG-S-2525; chitinase inhibitors such as OATD-01; mitogen-activated
protein kinase-activated protein kinase 2 (MAPKAPK2) inhibitors
such as MMI-0100; transforming growth factor beta 1 (TGF-beta1)
small interfering RNA such as TRK250/BNC-1021; or lysophosphatidic
acid receptor antagonists such as BMS-986278.
Anti-tussive agents are, for example, purinoceptor 3 (P2X3)
receptor antagonists such as gefapixant, S-600918, BAY-1817080, or
BLU-5937; neurokinin 1 (NK-1) receptor antagonist such as
Orvepitant, Aprepitant; nicotinic acetylcholine receptor alpha 7
subunit stimulator such as ATA-101/bradanicline; codeine,
gabapentin, pregablin, or azithromycin. Anti-inflammatory agents
are, for example, corticosteroids such as prednisolone or
dexamethasone; cyclo-oxygenase-2 (COX2) inhibitors such as
celecoxib, rofecoxib, parecoxib, valdecoxib, deracoxib, etoricoxib
or lumiracoxib; prostaglandin E2 antagonists; leukotriene B4
antagonists; leukotriene D4 antagonists such as monteleukast;
5-lipoxygenase inhibitors; or other nonsteroidal anti-inflammatory
agents (NSAIDs) such as aspirin, diclofenac, diflunisal, etodolac,
ibuprofen or indomethacin.
Anti-atopic dermatitis agents are, for example, cyclosporin,
methotrexate, mycophenolate mofetil, azathioprine,
phosphodiesterase inhibitors (e.g. apremilast, crisaborole), Janus
Associated Kinase (JAK) inhibitors (e.g. tofacitinib), neutralizing
antibodies against IL-4/IL13 (e.g. dupilamab), IL-13 (e.g.
lebrikizumab, tralokinumab) and IL-31 (nemolizumab). Analgesics
are, for example, of the opioid type, such as morphine,
oxymorphine, levopanol, oxycodon, propoxyphene, nalmefene,
fentanyl, hydrocondon, hydromorphone, meripidine, methadone,
nalorphine, naloxone, naltrexone, buprenorphine, butorphanol,
nalbuphine, pentazocine; or of the non-opioid type, such as
acetophenamine.
Anti-depressants are, for example, tricyclic anti-depressants such
as amitriptyline, clomipramine, despramine, doxepin, desipramine,
imipramine, nortriptyline; selective serotonin reuptake inhibitor
anti-depressants (SSRIs) such as fluoxetine, paroxetine,
sertraline, citalopram, escitalopram; norepinephrine reuptake
inhibitor anti-depressants (SNRIs) such as maprotiline,
lofepramine, mirtazapine, oxaprotiline, fezolamine, tomoxetine,
mianserin, buproprion, hydroxybuproprion, nomifensine, viloxazine;
dual serotonin-norepinephrine reuptake inhibitor anti-depressants
(SNRIs) such as duloxetine, venlafaxine, desvenlafaxine,
levomilnacipran; atypical antidepressants such as trazodone,
mirtazapine, vortioxetine, vilazodone, bupropion; or monoamine
oxidase inhibitor anti-depressantss (MAOIs) such as
tranylcypromine, phenelzine, or isocarboxazid.
Anxiolytics are, for example, benzodiazepines such as alprazolam,
bromazepam, chlordiazepoxide, clonazepam, clorazepate, diazepam,
flurazepam, lorazepam, oxazepam, temazepam, triazolam, or
tofisopam; or they are nonbenzodiazepine hypnoticssuch as
eszopiclone, zaleplon, zolpidem, or zopiclone; or they are
carbamates e.g. meprobamate, carisoprodol, tybamate, or lorbamate;
or they are antihistamines such as hydroxyzine, chlorpheniramine or
diphenhydramine.
Sedatives are, for example, barbiturate sedatives, such as
amobarbital, aprobarbital, butabarbital, butabital, mephobarbital,
metharbital, methohexital, pentobarbital, secobarbital, talbutal,
theamylal, or thiopental; or they are non-barbiturate sedatives
such as glutethimide, meprobamate, methaqualone or
dichloalphenazone.
Skeletal muscle relaxants are, for example, baclofen, meprobamate,
carisoprodol, cyclobenzaprine, metaxalone, methocarbamol,
tizanidine, chlorzoxazone or orphenadrine. Other suitable
combination partners are inhibitors of Acetylcholinesterase
inhibitors such as donepezil; 5-HT-3 anatgonists such as
ondansetron; metabotropic glutamate receptor antagonists;
antiarrhythmics such as mexiletine or phenytoin; or NMDA receptor
antagonists. Further suitable combination partners are incontinence
medications, for example, anticholinergics such as oxybutynin,
tolterodine, darifenacin, fesoterodine, solifenacin or trospium; or
they are bladder muscle relaxants such as mirabegron; or they are
alpha blockers such as tamsulosin, alfuzosin, silodosin, doxazosin
or terazosin.
The dosage for the combination partners mentioned above is usually
1/5 of the lowest dose normally recommended up to 1/1 of the
normally recommended dose. Therefore, in another aspect, this
invention relates to the use of a compound according to the
invention in combination with one or more additional therapeutic
agents described hereinbefore and hereinafter for the treatment of
diseases or conditions which may be affected or which are mediated
by TRPA1, in particular diseases or conditions as described
hereinbefore and hereinafter.
In a further aspect this invention relates to a method for treating
a disease or condition which can be influenced by the inhibition of
TRPA1 in a patient that includes the step of administering to the
patient in need of such treatment a therapeutically effective
amount of a compound of formula (I) or a pharmaceutically
acceptable salt thereof in combination with a therapeutically
effective amount of one or more additional therapeutic agents.
In a further aspect this invention relates to the use of a compound
of formula (I) or a pharmaceutically acceptable salt thereof in
combination with one or more additional therapeutic agents for the
treatment of diseases or conditions which can be influenced by the
inhibition of TRPA1 in a patient in need thereof.
In yet another aspect the present invention relates a method for
the treatment of a disease or condition mediated by TRPA1 activity
in a patient that includes the step of administering to the
patient, preferably a human, in need of such treatment a
therapeutically effective amount of a compound of the present
invention in combination with a therapeutically effective amount of
one or more additional therapeutic agents described in hereinbefore
and hereinafter.
The use of the compound according to the invention in combination
with the additional therapeutic agent may take place simultaneously
or at staggered times.
The compound according to the invention and the one or more
additional therapeutic agents may both be present together in one
formulation, for example a tablet or capsule, or separately in two
identical or different formulations, for example as a so-called
kit-of-parts.
Consequently, in another aspect, this invention relates to a
pharmaceutical composition that comprises a compound according to
the invention and one or more additional therapeutic agents
described hereinbefore and hereinafter, optionally together with
one or more inert carriers and/or diluents.
In yet another aspect the present invention relates to the use of a
compound according to the invention in a cough-measuring
device.
Other features and advantages of the present invention will become
apparent from the following more detailed examples which
illustrate, by way of example, the principles of the invention.
PREPARATION
The compounds according to the present invention and their
intermediates may be obtained using methods of synthesis which are
known to the one skilled in the art and described in the literature
of organic synthesis. Preferably, the compounds are obtained in
analogous fashion to the methods of preparation explained more
fully hereinafter, in particular as described in the experimental
section. In some cases, the order in carrying out the reaction
steps may be varied. Variants of the reaction methods that are
known to the one skilled in the art but not described in detail
here may also be used.
The general processes for preparing the compounds according to the
invention will become apparent to the one skilled in the art
studying the following schemes. Starting materials may be prepared
by methods that are described in the literature or herein, or may
be prepared in an analogous or similar manner. Any functional
groups in the starting materials or intermediates may be protected
using conventional protecting groups. These protecting groups may
be cleaved again at a suitable stage within the reaction sequence
using methods familiar to the one skilled in the art.
The compounds according to the invention are prepared by the
methods of synthesis described hereinafter in which the
substituents of the general formulae have the meanings given
hereinbefore. These methods are intended as an illustration of the
invention without restricting its subject matter and the scope of
the compounds claimed to these examples.
Where the preparation of starting compounds is not described, they
are commercially obtainable or may be prepared analogously to known
compounds or methods described herein. Substances described in the
literature are prepared according to the published methods of
synthesis.
Compounds of formula I may be prepared as shown in Scheme I
below.
##STR00022##
In scheme I, alpha-cyano ketones of substructure (A) are reduced in
an enantioselective fashion by using appropriate catalytic systems
using a transition metal complex (of e.g. Ru or Ir) in combination
with a chiral ligand (e.g.
[(1S,2S)-(-)-2-amino-1,2-diphenylethyl](4-toluenesulfonyl)amido).
To the corresponding cyano-alcohols (B) is added hydroxylamine to
create the dihydroxypropanimidamides (C). The ring-closure to the
chloromethylenoxadiazoles (D) can be achieved by stirring the
reaction mixture together with chloro acetyl chloride. Finally
compounds of general formula (I) can be prepared by combining the
compounds of general formula (D) together with the appropriate
thienopyrimidinones in the presence of a base (e.g.
K.sub.2CO.sub.3).
Tricyclic thienopyrimidone compounds may be prepared as shown in
Scheme II below.
##STR00023##
In scheme II, a suitable thieno-tetrahydropyridine precursor is
converted with an appropriate reagent such as formamidine,
formamide or a salt thereof in a suitable solvent (e.g. EtOH) at
elevated temperatures (e.g. 100.degree. C.) into the tricyclic
thieno-pyrimidone compound (C). The tetrahydropyridine core is then
oxidized using a suitable oxidant (e.g. KMnO4) and is accelerated
by the presence of a chelating reagent (e.g. 18-crown-6). This
reaction is typically performed in a non-polar solvent (e.g. DCM)
and run preferentially at ambient temperature. In case R3 resembles
a protecting group (e.g. BOC), this group can be removed using
suitable conditions for deprotection (e.g. TFA/DCM or HCl/dioxane
at RT for R3=BOC).
EXAMPLES
Preparation
The compounds according to the invention and their intermediates
may be obtained using methods of synthesis which are known to the
one skilled in the art and described in the literature of organic
synthesis for example using methods described in "Comprehensive
Organic Transformations", 2nd Edition, Richard C. Larock, John
Wiley & Sons, 2010, and "March's Advanced Organic Chemistry",
7th Edition, Michael B. Smith, John Wiley & Sons, 2013.
Preferably the compounds are obtained analogously to the methods of
preparation explained more fully hereinafter, in particular as
described in the experimental section. In some cases the sequence
adopted in carrying out the reaction schemes may be varied.
Variants
ofthesereactionsthatareknowntotheskilledartisanbutarenotdescribedindetail
herein may also be used. The general processes for preparing the
compounds according to the invention will become apparent to the
skilled man on studying the schemes that follow. Starting compounds
are commercially available or may be prepared by methods that are
described in the literature or herein, or may be prepared in an
analogous or similar manner. Before the reaction is carried out,
any corresponding functional groups in the starting compounds may
be protected using conventional protecting groups. These protecting
groups may be cleaved again at a suitable stage within the reaction
sequence using methods familiar to the skilled man and described in
the literature for example in "Protecting Groups", 3rd Edition,
Philip J. Kocienski, Thieme, 2005, and "Protective Groups in
Organic Synthesis", 4th Edition, Peter G. M. Wuts, Theodora W.
Greene, John Wiley & Sons, 2006. The terms "ambient
temperature" and "room temperature" are used interchangeably and
designate a temperature of about 20.degree. C., e.g. between 19 and
24.degree. C.
TABLE-US-00010 Abbreviations: ACN acetonitrile Aq. aqueous .degree.
C. Degree celsius CDI 1,1'-Carbonyldiimidazole CyH cyclohexane
conc. concentrated DCM dichloro methane DIPEA
N,N-diisopropylethylamine DMF N,N-dimethylformamide DMSO dimethyl
sulfoxide ESI-MS Electrospray ionisation mass spectrometry EtOAc
ethyl acetate ex example eq equivalent h hour HATU
N,N,N',N'-tetramethyl-O-(7-azabenzotriazol-1-yl)uranium
hexafluoro-phosphate HCl Hydrochloric acid HPLC High performance
liquid chromatography Int Intermediate L liter LiHMDS
Lithium-bis(trimethylsilyl)amid MeOH methanol NaHCO.sub.3 sodium
bicarbonate min minute mL milliliter MTBE tert-butylmethylether NaH
Sodium hydride Pd/C palladium on activated carbon PE petroleum
ether RT room temperature (about 20.degree. C.) sat. saturated TBTU
Benzotriazolyl tetramethyluronium tetrafluoroborate TEA
triethylamine TFA trifluoroacetic acid THF tetrahydrofuran TLC
Thin-layer chromatography on SiO2
Preparation of Starting Compounds
Intermediate I
Intermediate I.1 (General Route)
(3S)-3-(4-chlorophenyl)-3-hydroxypropanenitrile
##STR00024##
10.0 g (55.7 mmol) 4-Chlorobenzoylacetonitrile are added to 100 mL
ACN under inert atmosphere. 142 mg (0.23 mmol)
Chloro([(S,2S)-(-)-2-amino-1,2-diphenylethyl](4-toluenesulfonyl)amido)(me-
sitylene)ruthenium (II) (CAS 174813-81-1) are added before 8.30 mL
(19.8 mmol) formic acid triethylamine complex (5:2) are added
dropwise. After stirring at RT for 3 h the solvent is removed in
vacuo. To the remaining crude mixture is added water and this
mixture is extracted two times with EtOAc. The organic layers are
combined, dried over MgSO4 and the solvent is removed in vacuo.
C.sub.9H.sub.8ClNO (M=181.6 g/mol)
ESI-MS: 226 [M+HCOO].sup.-
R.sub.t(HPLC): 0.81 min (method B)
The following compounds are prepared according to the general
procedure (intermediateI.1) described above:
TABLE-US-00011 HPLC- retention time (method) Int. Starting
materials Structure ESI-MS [min] I.2 ##STR00025## ##STR00026##
EI-MS: 187 [M*].sup.+ 0.41 (A) I.3 ##STR00027## ##STR00028## 184 [M
+ Na].sup.+ 0.76 (B) I.4 ##STR00029## ##STR00030## 188 [M +
Na].sup.+ 0.72 (B) I.5 ##STR00031## ##STR00032## EI-MS: 183
[M*].sup.+ 0.79 (B) I.6 ##STR00033## ##STR00034## 184 [M + H].sup.+
0.76 (B) I.7 Example V ##STR00035## 250 [M + Na].sup.+ 0.84 (B) I.8
##STR00036## ##STR00037## 214 [M + Na].sup.+ 0.64 (B)
Intermediate II
Intermediate II.1 (General Route)
(3S)-3-(4-chlorophenyl)-N,3-dihydroxypropanimidamidl
##STR00038##
A mixture of 9.82 g (54.1 mmol)
(3S)-3-(4-chlorophenyl)-3-hydroxypropanenitrile (intermediate 1.1),
and 8.00 mL (136 mmol) hydroxylamine (50% in water) are added to
100 mL MeOH and strirred at 75.degree. C. for 1.5 h. After cooling
down to RT, all volatiles are removed in vacuo to yield the crude
product which is used without further purification.
C.sub.9H.sub.11ClN.sub.2O.sub.2(M=214.6 g/mol)
ESI-MS: 215 [M+H].sup.+
R.sub.t (HPLC): 0.60 min (method B)
The following compounds are prepared according to the general
procedure (intermediate II.1) described above:
TABLE-US-00012 HPLC Starting retention time Int. materials
Structure ESI-MS (method) [min] II.2 I.2 ##STR00039## 221 [M +
H].sup.+ 0.27 (A) II.3 I.3 ##STR00040## 195 [M + H].sup.+ 0.57 (B)
II.4 I.3 ##STR00041## 199 [M + H].sup.+ 0.43 (B) II.5 I.5
##STR00042## 217 [M + H].sup.+ 0.54 (B) II.6 I.6 ##STR00043## 217
[M + H].sup.+ 0.55 (B) II.7 I.7 ##STR00044## 261 [M + H].sup.+ 0.66
(C) II.8 I.8 ##STR00045## 225 [M + H].sup.+ 0.24 (B)
Intermediate III
Intermediate III.1 (General Route)
(1S)-2-[5-(chloromethyl)-1,2,4-oxadiazol-3-yl]-1-(4-chlorophenyl)ethan-1-o-
l
##STR00046##
To 11.2 g (52.4 mmol) of intermediate II in 55 mL NMP are added
10.0 mL (57.8 mmol) DIPEA. The mixture is cooled to 0.degree. C.
before 4.60 mL (57.7 mmol) chloroacetyl chloride dissolved in 5 mL
NMP are slowly added and the mixture is stirred at constant
temperature for 45 min. The mixture is then heated up to 95.degree.
C. and stirring is continued for 4 h. After cooling down to RT, 200
mL water are added and the resulting mixture is extracted three
times with EtOAc. The organic layers are combined, dried over
MgSO.sub.4, filtered and the solvent is removed in vacuo. The
residue is purified by column chromatography (silica gel; PE/EtOAc,
7/3).
C.sub.11H.sub.10Cl.sub.2N.sub.2O.sub.2 (M=273.1 g/mol)
ESI-MS: 271 [M-H].sup.-
R.sub.t (HPLC): 0.93 min (method B)
The following compounds are prepared according to the general
procedure (intermediate III.1) described above:
TABLE-US-00013 HPLC retention time Starting (method) Int. materials
Structure ESI-MS [min] III.2 II.2 ##STR00047## 301 [M + Na].sup.+
0.90 (B) III.3 II.3 ##STR00048## 235 [M + H - H.sub.2O].sup.+ 0.92
(C) III.4 II.4 ##STR00049## 255 [M - H].sup.- 0.86 (B) III.5 II.5
##STR00050## 273 [M - H].sup.- 0.93 (B) III.6 II.6 ##STR00051## 273
[M - H].sup.- 0.93 (B) III.7 II.7 ##STR00052## EI-MS: 318
[M*].sup.- 0.96 (G) III.8 II.8 ##STR00053## 281 [M - H].sup.- 0.82
(G)
Intermediate IV
Intermediate IV.1 (General Route)
N,5-dimethyl-4-oxo-3H,4H-thieno[2,3-d]pyrimidine-6-carboxamide
##STR00054##
To 0.25 g (1.19 mmol)
5-methyl-4-oxo-3H,4H-thieno[2,3-d]pyrimidine-6-carboxylic acid in
30 mL THF are added 331 .mu.L (2.38 mmol) TEA and 429 mg (1.19
mmol) TBTU. The mixture is stirred at RT for 30 min before 654
.mu.L (1.31 mmol) of a methylamine solution (c=2 mol/L in THF) is
added. Stirring is continued at ambient temperature for 16 h. The
solvent is removed in vacuo before water and DCM are added. The
organic layer is separated, dried over Na.sub.2SO.sub.4 and the
solvent is removed in vacuo. The crude product is purified by
column chromatography (silica gel; DCM/MeOH/NH.sub.3 90/10/1).
C.sub.9H.sub.9N.sub.3O.sub.2S (M=223.3 g/mol)
ESI-MS: 224 [M+H].sup.+
R.sub.t (HPLC): 0.62 min (method C)
The following compounds are prepared according to the general
procedure (example IV.1) described above:
TABLE-US-00014 HPLC retention time (method) Int. Starting materials
Structure ESI-MS [min] IV.2 ##STR00055## ##STR00056## 274 [M +
H].sup.+ 0.72 (C) IV.3 ##STR00057## ##STR00058## 282 [M + H].sup.+
0.68 (C)
Intermediate V
3-(2,2-Difluoro-2H-1,3-benzodioxol-5-yl)-3-oxopropanenitrile
##STR00059##
To 0.63 mL (12.0 mmol) acetonitrile in 5 mL THF are added 18.0 mL
potassium 2-methyl-2-butoxide (conc: 2 mol/L in THF; 36.1 mmol) and
the resulting mixture is stirred for a short period. Then 2.60 g
(12.0 mmol) methyl 2,2-difluoro-2H-1,3-benzodioxole-5-carboxylate
are added and stirring is continued at RT for 30 min. The reaction
is quenched by the addition of aq. HCl solution (conc.: 1 mol/L).
EtOAc is added and the org. layer is separated, washed with brine
and dried over Na.sub.2SO.sub.4. The solvent is removed in vacuo
and the crude product is purified by column chromatography (silica
gel; CyH/EtOAc 75/25.fwdarw.57/43) to give the desired product.
C.sub.10H.sub.5F.sub.2NO.sub.3 (M=225.2 g/mol)
ESI-MS: 224 [M-H].sup.-
R.sub.t (HPLC): 0.89 min (method B)
Intermediate VI
tert-butyl
4,8-dioxo-5,6-dihydro-3H-pyrido[2,3]thieno[2,4-c]pyrimidine-7-c-
arboxylate
##STR00060##
To 500 mg (1.63 mmol) of tert-butyl
4-oxo-3,5,6,8-tetrahydropyrido[2,3]thieno[2,4-c]pyrimidine-7-carboxylate
in 8.0 mL of DCM is added 86.0 mg (0.325 mmol) of 18-crown-6 and
514 mg (3.25 mmol) of potassium permanganate. The reaction mixture
is stirred at ambient temperature overnight. The reaction mixture
is then quenched by addition of methanol and aqueous 10%
Na.sub.2S203 solution. The precipitated solid is removed by
filtration and the filtrate is concentrated. The residue is
dissolved in methanol and DMF, filtered and purified by preparative
HPLC (ACN/H.sub.2O/TFA gradient). The product containing fractions
are combined and the acetonitrile is removed under reduced
pressure. The remaining aqueous product solution is extracted with
DCM (2.times.). The combined organic extracts are dried over sodium
sulfate to give the desired product.
C.sub.14H.sub.15N.sub.3O.sub.4S (M=321.3 g/mol)
ESI-MS: 322 [M+H].sup.30
R.sub.t (HPLC): 0.46 min (method A)
Intermediate VII
3,5,6,7-tetrahydropyrido[2,3]thieno[2,4-c]pyrimidine-4,8-dione
##STR00061##
To 264 mg (0.822 mmol) of tert-butyl
4,8-dioxo-5,6-dihydro-3H-pyrido[2,3]thieno[2,4-c]pyrimidine-7-carboxylate
in 6 mL DCM is added 380 .mu.L (4.926 mmol) trifluoroacetic acid.
The resulting mixture is stirred at ambient temperature for 45 min.
All volatiles are removed in vacuo and the residue is used as
obtained in the next step.
C.sub.9H.sub.7N.sub.3O.sub.2S (M=221.2 g/mol)
ESI-MS: 222 [M+H].sup.+
R.sub.t (HPLC): 0.22 min (method A)
Intermediate VIII
Intermediate VIII.1 (General Route)
##STR00062##
To 5.00 g (26.8 mmol) 2-amino-4-chloro-3-cyano-5-formylthiophene
(CAS: 104366-23-6) in 25 mL pyridine is added 5.70 mL (42.9 mmol)
N,N-dimethylformamide dimethyl acetal. The mixture is stirred at
100.degree. C. for 3 h. After cooling to ambient temperature, the
mixture is concentrated under reduced pressure. The residue is
resuspended in DCM and washed with water. The organic phase is
dried over Na.sub.2SO.sub.4 and concentrated to yield the desired
product.
C.sub.9H.sub.8ClN.sub.3OS (M=241.7 g/mol)
ESI-MS: 242 [M+H].sup.+
R.sub.t (HPLC): 1.04 min (method B)
Intermediate IX
Intermediate IX.1 (General Route)
##STR00063##
To 1.00 g (4.14 mmol) of intermediate IX.1 in 10 mL formic acid is
added 678 mg (8.27 mmol) sodium acetate. The reaction mixture is
stirred at reflux overnight. After cooling to ambient temperature,
the mixture is poured onto ice-cold water. It is then diluted with
dichloromethane and concentrated to dryness. The residue is
suspended in dichloromethane and the remaining salts are filtered
off. The filtrate is concentrated to yield the desired product.
C.sub.7H.sub.3ClN.sub.2O.sub.2S (M=214.6 g/mol)
ESI-MS: 213 [M-H].sup.-
R.sub.t (HPLC): 0.87 min (method B)
Intermediate X
Intermediate X.1 (General Route)
##STR00064##
To 150 mg (699 .mu.mol) of intermediate IX.1 in 4.0 mL DMF is added
473 mg (769 .mu.mol) potassium peroxymonosulfate. The reaction
mixture is added at ambient temperature for 18 h. The mixture is
then purified by preparative HPLC (H.sub.2O/ACN/TFA) to yield the
desired product.
C.sub.7H.sub.3ClN.sub.2O.sub.3S (M=230.6 g/mol)
ESI-MS: 229 [M-H].sup.-
R.sub.t (HPLC): 0.58 min (method C)
Intermediate XI
Intermediate XI.1 (General Route)
##STR00065##
To 45.0 mg (0.195 mmol) of example X.1 in 1.0 mL DMF is added 81.7
mg (0.215 mmol)
1-[Bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium
3-oxid hexafluorophosphate, 74.8 .mu.L (0.429 mmol)
diisopropylethylamine and 1.17 mL (0.5 M, 0.585 mmol) of a solution
of ammonia in THF. The reaction mixture is stirred at ambient
temperature for 18 h. The reaction mixture is then purified by
preparative HPLC (H.sub.2O/ACN/NH.sub.3) to yield the desired
product.
C.sub.7H.sub.4ClN.sub.3O.sub.2S (M=229.6 g/mol)
ESI-MS: 230 [M+H].sup.+
R.sub.t (HPLC): 0.52 min (method C)
Preparation of Final Compounds
Example 1
Example 1 (General Route)
3-({3-[(2S)-2-(4-chlorophenyl)-2-hydroxyethyl]-1,2,4-oxadiazol-5-yl}methyl-
)-5-methyl-4-oxo-3H,4H-thieno[2,3-d]pyrimidine-6-carboxamide
##STR00066##
2.00 g (9.56 mmol) of
5-Methyl-4-oxo-3H,4H-thieno[2,3-d]pyrimidin-6-carboxamide are added
to 20 mL DMA before 4.30 g (31.1 mmol) K.sub.2CO.sub.3 are added.
The mixture is stirred at RT for 20 minutes before 2.70 g (9.89
mmol) of example III.1 in 5 mL DMA are added. Strirring is
continued at 50.degree. C. for 3 h. After cooling down to RT, the
mixture is filtered and purified by HPLC (ACN/H.sub.2O gradient
containing 0.3% TFA) to yield the desired product.
C.sub.19H.sub.16ClN.sub.5O.sub.4S (M=445.9 g/mol)
ESI-MS: 446 [M+H].sup.+
R.sub.t (HPLC): 0.82 min (method B)
.sup.1H NMR (400 MHz, DMSO-d6) .delta. ppm: 2.70 (s, 3H), 2.92-3.05
(m, 2H), 4.94 (dt, J=7.60, 5.30 Hz, 1H), 5.52 (s, 2H), 5.60 (d,
J=4.80 Hz, 1H), 7.26-7.39 (m, 4H), 7.69 (br s, 2H), 8.63 (s,
1H).
The following compounds are prepared according to the general
procedure (example 1) described above:
TABLE-US-00015 Reaction Ex. Starting materials Structure conditions
2 III.3 + 5-Methyl-4-oxo- 3H,4H-thieno[2,3- d]pyrimidin-6-
carboxamide ##STR00067## Solvent: DMF 3 III.2 + 5-Methyl-4-oxo-
3H,4H-thieno[2,3- d]pyrimidin-6- carboxamide ##STR00068## Reaction
done at RT 4 III.4 + 5-Methyl-4-oxo- 3H,4H-thieno[2,3-
d]pyrimidin-6- carboxamide ##STR00069## Reaction done at RT 5 III.5
+ 5-Methyl-4-oxo- 3H,4H-thieno[2,3- d]pyrimidin-6- carboxamide
##STR00070## Reaction done at RT 6 III.6 + 5-Methyl-4-oxo-
3H,4H-thieno[2,3- d]pyrimidin-6- carboxamide ##STR00071## Reaction
done at RT 7 III.7 + 5-Methyl-4-oxo- 3H,4H-thieno[2,3-
d]pyrimidin-6- carboxamide ##STR00072## RT, 16 h 8 III.8 +
5-Methyl-4-oxo- 3H,4H-thieno[2,3- d]pyrimidin-6- carboxamide
##STR00073## 60.degree. C., 1 h 9 III.8 + IV.1 ##STR00074## RT, DMF
10 III.4 + IV.1 ##STR00075## RT, DMF 11 III.1 + IV.1 ##STR00076##
50.degree. C., 0.5 h, DMF 12 III.1 + IV.2 ##STR00077## 50.degree.
C., 0.5 h, DMF 13 III.1 + IV.3 ##STR00078## 50.degree. C., 0.5 h,
DMF 14 III.5 + IV.1 ##STR00079## RT, DMF 15 III.1 + VI.1
##STR00080## RT, DMF 16 III.1 + XI.1 ##STR00081## RT, DMF, 18 h
Analytic data for the final compounds described in the table
above:
TABLE-US-00016 HPLC re- tention time (method) Ex. ESI-MS [min]
.sup.1H NMR (400 MHz, DMSO-d6) .delta. ppm 2 426 0.56 2.25 (s, 3H),
2.70 (s, 3H), 2.91 (dd, J = 14.6, 5.5 Hz, 1H), [M + H].sup.+ (D)
2.99 (dd, J = 14.7, 8.1 Hz, 1H), 4.86 (td, J = 7.98, 5.3 Hz, 1H),
5.41 (d, J = 4.8, 1H), 5.51 (s, 2H), 7.04-7.11 (m, 2H), 7.17-7.23
(m, 2H), 7.69 (s, 2H), 8.64 (s, 1H) 3 452 0.81 2.69 (s, 3H), 3.19
(dd, J = 14.8, 8.1 Hz, 1H), 3.25 (dd, [M + H].sup.+ (B) J = 15.1,
5.8 Hz, 1H), 5.05-5.12 (m, 1H), 5.52 (s, 2H), 5.91 (d, J = 5.7 Hz,
1H), 6.75 (s, 1H), 7.23 (t, J = 7.4 Hz, 1H), 7.26 (t, J = 7.5 Hz,
1H), 7.51 (d, J = 8.1 Hz, 1H), 7.55 (d, J = 7.5 Hz, 1H), 7.69 (br
s, 2H), 8.62 (s, 1H) 4 430 0.82 2.70 (s, 3H), 2.95 (dd, J = 14.6,
5.6 Hz, 1H), 3.01 (dd, [M + H].sup.+ (B) J = 14.6, 8.2 Hz, 1H),
4.94 (dd, J = 8.1, 5.4 Hz, 1H), 5.52 (s, 2H), 7.06-7.12 (m, 2H),
7.34-7.40 (m, 2H), 7.69 (s, 2H), 8.64 (s, 1H) 5 448 0.55 2.70 (s,
3H), 3.00 (d, J = 6.6 Hz, 2H), 4.95 (t, J = 6.7 Hz, [M + H].sup.+
(E) 1H), 5.52 (s, 2H), 5.54-5.85 (m, 1H), 7.14-7.21 (m, 1H), 7.31
(dt, J = 10.7, 8.4 Hz, 1H), 7.39 (ddd, J = 11.9, 8.0, 1.8 Hz, 1H),
7.69 (br s, 2H), 8.63 (s, 1H) 6 448 0.63 2.69 (s, 3H), 2.99 (dd, J
= 14.6, 8.0 Hz, 1H), 3.04 (dd, [M + H].sup.+ (F) J = 14.6, 5.3 Hz,
1H), 4.97 (br t, J = 6.1 Hz, 1H), 5.52 (s, 2H), 5.75 (br s, 1H),
7.01-7.10 (m, 3H), 7.69 (s, 2H), 8.63 (s, 1H) 7 492 0.71 2.70 (s,
3H), 2.95-3.06 (m, 2H), 4.97 (dt, J = 7 .7 , 5.4 Hz, [M + H].sup.+
(H) 1H), 5.52 (s, 2H), 5.66 (d, J = 4.8 Hz, 1H), 7.16 (dd, J = 8.3,
1.5 Hz, 1H), 7.28 (d, J = 8.2 Hz, 1H), 7.40 (d, J = 1.5 Hz, 1H),
7.69 (br s, 2H), 8.63 (s, 1H) 8 456 0.52 2.70 (s, 3H), 2.91 (dd, J
= 14.6, 5.3 Hz, 1H), 2.98 (dd, [M + H].sup.+ (I) J = 14.6, 8.4 Hz,
1H), 4.85 (dd, J = 8.2, 5.4 Hz, 1H), 5.43 (br s, 1H), 5.52 (s, 2H),
5.96 (dd, J = 3.5, 1.0 Hz, 2H), 6.75-6.80 (m, 2H), 6.92 (s, 1H),
7.69 (br s, 2H), 8.64 (s, 1H) 9 470 0.60 2.67 (s, 3H), 2.78 (d, J =
4.6 Hz, 3H), 2.91 (dd, J = 14.4, [M + H].sup.+ (H) 5.3 Hz, 1H),
2.98 (dd, J = 14.6, 8.4 Hz, 1H), 4.85 (dd, J = 8.4, 5.3 Hz, 1H),
5.51 (s, 2H), 5.96 (dd, J = 3.3, 0.9 Hz, 2H), 6.77 (s, 2H), 6.92
(s, 1H), 8.18 (br q, J = 4.6 Hz, 1H), 8.64 (s, 1H) 10 444 0.64 2.67
(s, 3H), 2.78 (d, J = 4.6 Hz, 3H), 2.95 (dd, J = 14.6, [M +
H].sup.+ (H) 5.6 Hz, 1H), 3.01 (dd, J = 14.6, 8.1 Hz, 1H), 4.94
(dd, J = 8.0, 5.5 Hz, 1H), 5.51 (s, 2H), 7.04 - 7.12 (m, 2H),
7.33-7.39 (m, 2H), 8.18 (q, J = 4.5 Hz, 1H), 8.64 (s, 1H) 11 460
0.92 2.68 (s, 3H), 2.78 (d, J = 4.6 Hz, 3H), 2.92-3.04 (m, 2H), [M
+ H].sup.+ (C) 4.94 (dd, J = 7.8, 5.8 Hz, 1H), 5.52 (s, 2H),
7.30-7.36 (m, 4H), 8.18 (q, J = 4.4 Hz, 1H), 8.63 (s, 1H) 12 510
0.98 2.69 (s, 3H), 2.92-3.05 (m, 2H), 3.67 (tdd, J = 15.7, 5.7, [M
+ H].sup.+ (C) 4.1 Hz, 2H), 4.94 (dt, J = 7.6, 5.3 Hz, 1H), 5.52
(s, 2H), 5.60 (d, J = 4.8 Hz, 1H), 6.15 (tt, J = 55.9, 3.8 Hz, 1H),
7.26-7.41 (m, 4H), 8.61-8.68 (m, 2H) 13 518 0.93 1.12 (s, 6H), 2.69
(s, 3H), 2.92-3.05 (m, 2H), 3.24 (br d, [M + H].sup.+ (C) J = 6.1
Hz, 2H), 4.54 (s, 1H), 4.90-4.98 (m, 1H), 5.52 (s, 2H), 5.60 (d, J
= 4.8 Hz, 1H), 7.29-7.37 (m, 4H), 8.03 (t, J = 6.1 Hz, 1H), 8.64
(s, 1H) 14 462 0.68 2.67 (s, 3H), 2.78 (d, J = 4.6 Hz, 3H),
2.96-2.04 (m, 2H), [M + H].sup.+ (H) 4.92-4.98 (m, 1H), 5.51 (s,
2H), 5.67 (br d, J = 4.2 Hz, 1H), 7.14-7.21 (m, 1H), 7.31 (dt, J =
10.8, 8.4 Hz, 1H), 7.39 (ddd, J = 11.8, 8.0, 2.0 Hz, 1H), 8.18 (q,
J = 4.5 Hz, 1H), 8.63 (s, 1H) 15 458 0.46 2.96 (dd, J = 14.4, 5.6
Hz, 1H), 3.01 (dd, J = 14.6, 7.9 Hz, [M + H].sup.+ (A) 1H), 3.13
(t, J = 7.0 Hz, 2H), 3.48-3.53 (m, 2H), 4.94 (t, J = 6.7 Hz, 1H),
5.54 (s, 2H), 5.60 (br s, 1H), 7.30-7.37 (m, 4H), 7.95-8.00 (m,
1H), 8.67 (s, 1H) 16 466 0.85 2.93-3.04 (m, 2H), 3.17 (d, J = 5.2
Hz, 1H), 4.94 (dt, [M + H].sup.+ (C) J = 7.6, 5.4 Hz, 1H), 5.53 (s,
2H), 5.60 (d, J = 4.8 Hz, 1H), 7.30-7.37 (m, 4H), 7.70 (hr s, 1H),
8.15 (hr s, 1H), 8.73 (s, 1H)
Analytical HPLC Methods Method A
TABLE-US-00017 Vol % water Flow time (min) (incl. 0.1% TFA) Vol %
ACN [mL/min] 0.00 99 1 1.6 0.02 99 1 1.6 1.00 0 100 1.6 1.10 0 100
1.6
Analytical column: XBridge BEH C18_2.1.times.30 mm, 1.7 .mu.m;
column temperature: 60.degree. C.
Method B
TABLE-US-00018 Vol % water Flow time (min) (incl. 0.1% TFA) Vol %
ACN [mL/min] 0.00 97 3 2.2 0.20 97 3 2.2 1.20 0 100 2.2 1.25 0 100
3.0 1.40 0 100 3.0
Analytical column: Stable Bond (Agilent) 1.8p m; 3.0.times.30 mm;
column temp: 60.degree. C.
Method C
TABLE-US-00019 Vol % water Flow time (min) (incl. 0.1% TFA) Vol %
ACN [mL/min] 0.00 97 3 2.2 0.20 97 3 2.2 1.20 0 100 2.2 1.25 0 100
3.0 1.40 0 100 3.0
Analytical column: Sunfire (Waters) 2.5 m; 3.0.times.30 mm; column
temperature: 60.degree. C.
Method D
TABLE-US-00020 Gradient/Solvent Vol % water (incl. 0.1% Vol % Time
[min] NH.sub.3) Acetonitrile Flow [ml/min] 0.0 95.0 5.0 1.5 1.3 0.0
100.0 1.5 1.5 0.0 100.0 1.5 1.6 95.0 5.0 1.5
Preparative column: XBridge (Waters) C18_3.0.times.30 mm_2.5 .mu.m;
column temperature: 60.degree. C.
Method E
TABLE-US-00021 Gradient/Solvent Vol % water (incl. Vol % Time [min]
0.1% NH.sub.3) Acetonitrile Flow [ml/min] 0.0 95.0 5.0 1.5 1.3 0.0
100.0 1.5 1.5 0.0 100.0 1.5 1.6 95.0 5.0 1.5
XBridge C18_3.0.times.30 mm 2.5 .mu.m (Waters); column temperature:
60.degree. C.
Method F
TABLE-US-00022 Gradient/Solvent Vol % water Vol.- % ACN Flow Time
[min] (incl. 0.1% TFA) (incl. 0.08% TFA) [ml/min] 0.0 95.0 5.0 1.5
1.3 0.0 100.0 1.5 1.5 0.0 100.0 1.5 1.6 95.0 5.0 1.5
Preparative column: Sunfire (Waters) C18_3.0.times.30 mm_2.5 .mu.m;
column temperature: 60.degree. C.
Method G
TABLE-US-00023 Vol % water Flow time (min) (incl. 0.1% NH.sub.4OH)
Vol % ACN [mL/min] 0.00 97 3 2.2 0.20 97 3 2.2 1.20 0 100 2.2 1.25
0 100 3 1.40 0 100 3
Analytical column: XBridge C18 (Waters) 2.5 .mu.m; 3.0.times.30 mm;
column temperature: 60.degree. C.
Method H
TABLE-US-00024 Vol % water Vol.- % ACN Flow time (min) (incl. 0.1%
TFA) (incl. 0.08 % TFA) [mL/min] 0.00 95 5 1.5 1.3 0 100 1.5 1.5 0
100 1.5 1.6 95 5 1.5
Analytical column: Sunfire C18 (Waters) 2.5 .mu.m; 3.0.times.30 mm;
column temperature: 60.degree. C.
Method I
TABLE-US-00025 Gradient/Solvent Vol % water Time [min] (incl. 0.1%
TFA) Vol % ACN Flow [ml/min] 0.0 50.0 50.0 1.5 0.02 50.0 50.0 1.5
1.0 0.0 100.0 1.5 1.1 0.0 100.0 1.5
Analytical column: Sunfire (Waters) C18_2.1.times.30 mm_2.5 .mu.m;
column temperature: 60.degree. C.
* * * * *
References